Jundishapur Journal of Microbiology
Volume:10 Issue: 8, Aug 2017

Microorganisms represent a potentially valuable source of biologically significant compounds that should be explored for their potential agrochemical use.
The compounds ZM-1 and ZM-2 were isolated from the extract of the fermentation broth of Streptomyces parvus 33 by bioassay-guided fractionation.
The compound ZM-1 was isolated in the form of a clear single crystal for the first time, and identified as holomycin via X-ray crystallography. The compound ZM-2 was characterized by IR, 1H-NMR, 13C-NMR, DEPT (90° and 135°), and MS analyses.
The compound ZM-1 showed a strong antibacterial activity against the tested bacteria, and its value of minimum inhibitory concentration (MIC) was greater than that of the positive control Ampicillin. The compound ZM-1 also exhibited significant antimicrobial activities against some plant pathogenic fungi. Nevertheless, ZM-2 showed no activities against the tested bacteria and plant pathogenic fungi.
The compound ZM-1 was isolated in the form of a clear single crystal for the first time, and identified as holomycin via X-ray crystallography. Streptomyces parvus 33 is a newly discovered producer of holomycin. ZM-1 showed a strong antibacterial activity against the tested bacteria. Therefore, the compound ZM-1 is a valuable lead compound for the development of agricultural fungicides while acts against bacteria, as well.

Sarcocystis species are obligatory intracellular parasites of many vertebrate hosts. Some pathogen species cause major economic loss and hygienic problems in the animal and human population, respectively.
The goal of the current study was conducted to identify Sarcocystis species in meat-producer animals and to evaluate the risk of transmission of parasites after consumption of infected meat by humans.
Fifty samples of sheep and cattle muscles were collected from the abattoir. The samples were collected from the heart, tongue, diaphragm, and skeletal muscles. The PCR method was used for amplifying the 18S ribosomal RNA gene for distinguish Sarcocystis species using 2 primers and 3 restricted enzymes including Hinf, Mbo1, and EcoR1.
The results showed that all cattle samples were infected by Sarcocystiscruzi (100%) and sheep samples were contaminated by S. tenella (80%) as well as S. capracanis (20%). No human Sarcocystis species were detected.
Meat-producer animals are infected by S. cruzi as well as S. tenella and the consumption of infected meat is not important for human sarcocystosis in this area.

Vulvovaginal candidiasis (VVC) is a common infection, affecting up to 75% of women at least once during their lifetime. In addition, approximately 5% of patients may experience recurrent VVC. Candida albicans is the most common causative agent of VVC. Overall, precise identification of the causative agents of VVC is necessary for effective treatment.
The purpose of this study was to identify the molecular characteristics and antifungal susceptibility of Candida species, isolated from women with VVC in cities of Shoush, Dezful, and Andimeshk, Khuzestan Province, Iran.
In this descriptive cross sectional study, vaginal samples were collected from 173 women with VVC, referred to gynecologists. The samples were cultured on Sabouraud dextrose agar, containing chloramphenicol. The ITS1-ITS4 region was amplified via polymerase chain reaction (PCR) assay and digested by MspI restriction enzyme. Antifungal susceptibility test was performed for 4 antifungal drugs (fluconazole, nystatin, itraconazole, and clotrimazole) via disk diffusion method.
Out of 173 patients, 95 (54.9%) showed VVC and 26 (27.4%) had recurrent VVC. The most common Candida species were C. albicans (70.5%), C. glabrata (20%), C. tropicalis (7.4%), and C. parapsilosis (2.1%), respectively. The antifungal susceptibility test showed resistance to fluconazole in 1 C. tropicalis, 2 C. albicans, and 3 C. glabrata isolates, while resistance to clotrimazole was detected in 1 C. albicans and 1 C. glabrata isolate.
According to the results of this study, approximately 30% of VVC infections were caused by non-C. albicans species, which should be considered by gynecologists due to their azole resistance.

Microorganisms play important roles in the macrobenthic bioremediation process. At present, studies regarding microeukaryotes in intertidal sediments are not enough, and studies of the microeukaryotic dynamics in the bioremediation process using macrobenthos have received much less attention.
This study aimed to reveal the microeukaryotic community structure and dynamics during the bioremediation process using macrobenthos in the intertidal sediments of Sansha Bay of China.
Twenty sediment samples were collected before and after bioremediation respectively. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as well as 18S rDNA clone libraries analyses were used.
The microeukaryotic communitywere divided into 10 phyla, in which Annelida and Arthropoda were the most dominant, and Gastrotricha and Nematoda were the second dominant groups. A small amount of Cercozoa, Bacillariophyta, Chlorophyta, Alveolata, Amoebazoa, Chytridiomycota, and some unknown eukaryotes were also present. The microeukaryotic community structure presented a clear variation with time. In June, Annelida was the most dominant, and Arthropoda, Nematoda, and Cercozoa were the subdominant phyla. While in September, phylum Annelida decreased dramatically, and Arthropoda and Gastrotricha increased greatly. December harbored a quite different and highest diversity of microeukaryotic community: the phylum Annelida, which was dominant in June and September, was not detected, while the phyla Bacillariophyta, Nematoda, and Alveolata increased greatly, and Amoebozoa and Chytridiomycota were only detected in December.
The bioremediation using macrobenthos changed the microeukartotic community structure and increased the diversity. The phyla Annelida and Arthropoda increased, while Gastrotricha, Cercozoa, and Alveolata decreased during the bioremediation.
The microeukaryotic community in the intertidal sediment of Sansha Bay of China, were divided into 10 phyla, and presented a clear variation with time. The bioremediation using macrobenthos changed the microeukartotic community structure and increased the diversity.

Rabies is a zoonotic infectious disease that infects the human and animal central nervous system. Worldwide, especially in low-income countries, this disease is still a burden for public health. Among the rabies virus proteins, phosphoprotein plays a very important role in viral infection, and this research found that immunization of rabies virus vaccines could widely induce antibody responses against phosphoprotein, therefore rabies virus phosphoprotein may be a useful target for development of rapid and low cost serological diagnosis test, and therapeutic drugs.
The aim of this study was to prepare anti-rabies virus phosphoprotein monoclonal antibody, which is used for rapid detection and diagnosis of rabies virus infection.
The phosphoprotein gene was amplified by the polymerase chain reaction (PCR), and sub-cloned in a his-tagged prokaryotic expression vector to generate a recombinant plasmid named pET-32a-RABV-P. Recombinant RABV-P was induced by isopropyl-β-D-Thiogalactopyranoside (IPTG), and then purified by the Ni-NTA purification system. Immunization of BALB/c mice with the recombinant protein was performed, and the spleen cells of the immunized mice and SP2/0 myeloma cells were fused together to obtain the monoclonal cell strains, and then identification of the characteristics of the antibody by the enzyme linked immunosorbent assay (ELISA), western blotting, and indirect immunofluorescence assay was done.
The prokaryotic expression vector of pET-32a-RABV-P was successfully constructed. The fusion protein was expressed in Escherichia coli Rosetta and purified. After cell fusion, a hybridoma cell line 1A4 was successfully obtained. The antibody titer of the anti-RABV-P ascites reached 256,000. The results of western blotting and indirect immunofluorescence assay indicated that 1A4 hybridoma cell line was able to produce specific antibodies against rabies virus phosphoprotein.
Recombinant rabies virus phosphoprotein could be successfully expressed in E. coli. A specific monoclonal antibody against rabies virus phosphoprotein has been successfully prepared.

Influenza is a major cause of morbidity and mortality worldwide. Each year, influenza viruses cause epidemics by evading pre-existing immunity through mutations in major surface glycoprotein hemagglutinin, which helps in attachment of the viral strain on the host cell surface. Due to high mutation rate, only currently circulating strains should be used in the vaccines.
The present study aimed at analyzing a dataset of complete amino acid sequences of HA to assess the extent of diversity among circulating strains of Iran, during years 2006 to 2013, and studying important amino acid changes as well as changes in predicted ligand binding sites that could enhance viral performance.
110 sequences from 17 provinces were downloaded, edited, and classified. The alignment of sequences and creation of phylogenetic trees and similarity matrices were done using bioinformatics software, such as MEGA6.0, BioEdit, DNAsisMAX, and DNAstar. Web-based analyses including SWISS- MODEL, Phyre2, and 3DLigandSite were used for evaluation of the second and third protein structures and prediction of ligand binding sites.
The results showed that 2009 was an important transition year, which classified the selected isolates into two different distinct groups. This shows the importance of changes made during possible mutations in the genomic structure of the virus, which have made it antigenically different from the previous years. This pandemic strain became dominant in the next years, and has been used as a standard vaccine strain from 2010 onwards.
The results of this study can shed further light on better understanding of the antigenic evolution of H1N1 influenza viruses and can be useful for epidemiological studies.

Infections with 5-Fluorocytosine (5-FC)-resistant Candida albicans isolates in China have rarely been reported in clinical settings. Here, we present two 5-FC-resistant C. albicans (Ca5508 and CaBD4291) strains, separately isolated from two patients in China.
The main aim of this study was to characterize the key genetic mutations responsible for 5-FC resistance of the two drug-resistant isolates from the first affiliated hospital of Nanchang university.
Multiplex PCR and phenotypic tests were used to confirm the isolates Ca5508 and CaBD4291. The standard micro broth dilution method M27-A3 was used for susceptibility testing of the two isolates. Molecular typing and analysis were performed by the combination of random amplified polymorphic DNA (RAPD), multilocus sequence typing (MLST), amplification and Sanger sequencing of 5-FC resistance-associated genes (FUR1 and FCA1).
The two isolates Ca5508 and CaBD4291 were identified as C. albicans by multiplex-PCR and phenotypic tests. The antifungal susceptibility testing showed that both of the isolates were resistant to 5-FC (MIC > 64µg/mL); Ca5508 was also resistant to fluconazole, ketoconazole, and itraconazole whereas CaBD4291 was not. The RAPD analysis demonstrated that the two isolates belonged to the same genotype. MLST typing identified two different sequence types: isolate CaBD4191 for ST732 and isolate Ca5508 for ST2975 (a new ST). Although the two STs were generally related, ST732 differed from ST2975 with 3/7 loci (AAC1, SYA1, VPS13). Surprisingly, the two isolates represented completely identical sequences of the two drug-associated genes FUR1 and FCA1. Compared to the susceptible reference strain SC5314, Ca5508 and CaBD4291 were found to have no mutation in FUR1; however, three missense mutations at positions 31, 83, and 107 of FCA1 were identified.
Here, we newly reported two 5-FC-resistant clinical isolates of C. albicans that represented the same three mutations in FCA1 gene contributed to 5-FC resistance of C. albicans.

Leukemia patients can easily become hypoimmunity after hemopoietic stem cell transplantation, endogenous infection often happens in these patients. Meanwhile, Klebsiella pneumoniae is often isolated from various specimens in inpatient. It can lead to infections in the whole body, especially in those immunosuppressed patients.
A 22-year-old girl with complete remission after chemotherapy for her acute lymphoblastic leukemia was presented to the 2nd affiliated hospital of Zhejiang University for hemopoietic stem cell transplantation. She got a good check-up, however, her spirit was a little bad. Her primitive lymphocyte (0.06%) and total lymphocyte (27.05%) in the peripheral blood were in the normal range. She received a 224 mL stem cell transfusion including mononuclear cell (4.87 × 108/kg) CD34 (0.6%) as well as reached the transplantation threshold of CD34 cell (3 × 106/kg). Nine days after transplantation, there were no severe side effects except a little bit of vomit. However, the symptom of diarrhea appeared first. K. pneumoniae was isolated from the stool, then invaded into the blood, and caused sepsis. It disseminated and caused multi-sites infection.
It should be kept in mind that K. pneumoniae can translocate across the intestinal epithelium. It is important to pay attention to the bacterium isolated from the intestine in immunosuppressed patients.